8AQH

NanoLuc-Y94A luciferase mutant

8AQH の概要

• エントリーDOI: 10.2210/pdb8aqh/pdb
• 分子名称: NanoLuc luciferase (2 entities in total)
• 機能のキーワード: luciferase, nanoluc, nluc, luciferin, furimazine, luminescent protein
• 由来する生物種: Oplophorus gracilirostris
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 40594.29

構造登録者

Nemergut, M.,Marek, M. (登録日: 2022-08-12, 公開日: 2023-08-23, 最終更新日: 2026-03-04)

主引用文献

Nemergut, M.,Pluskal, D.,Horackova, J.,Sustrova, T.,Tulis, J.,Barta, T.,Baatallah, R.,Gagnot, G.,Novakova, V.,Majerova, M.,Sedlackova, K.,Marques, S.M.,Toul, M.,Damborsky, J.,Prokop, Z.,Bednar, D.,Janin, Y.L.,Marek, M.
Illuminating the mechanism and allosteric behavior of NanoLuc luciferase.
Nat Commun, 14:7864-7864, 2023

PubMed Abstract: NanoLuc, a superior β-barrel fold luciferase, was engineered 10 years ago but the nature of its catalysis remains puzzling. Here experimental and computational techniques are combined, revealing that imidazopyrazinone luciferins bind to an intra-barrel catalytic site but also to an allosteric site shaped on the enzyme surface. Structurally, binding to the allosteric site prevents simultaneous binding to the catalytic site, and vice versa, through concerted conformational changes. We demonstrate that restructuration of the allosteric site can boost the luminescent reaction in the remote active site. Mechanistically, an intra-barrel arginine coordinates the imidazopyrazinone component of luciferin, which reacts with O via a radical charge-transfer mechanism, and then it also protonates the resulting excited amide product to form a light-emitting neutral species. Concomitantly, an aspartate, supported by two tyrosines, fine-tunes the blue color emitter to secure a high emission intensity. This information is critical to engineering the next-generation of ultrasensitive bioluminescent reporters.

PubMed: 38030625
DOI: 10.1038/s41467-023-43403-y

実験手法

X-RAY DIFFRACTION (2.798 Å)